CAJ | 학술논문

目的研究骨形态发生蛋白融合基因-4/7(BMP-4/7)和碱性成纤维细胞生长因子(bFGF)联合应用对体外培养兔骨髓基质干细胞(BMSCs)增殖和成骨活性的影响.方法体外培养BMSCs,在第3代细胞培养液中加入不同浓度的BMP-4/7和bFGF,依据加入不同基因浓度组合的不同分为5个实验组(A组:80 ng/mL BMP-4/7,B组:80 ng/mL bFGF,C组:30 ng/mL BMP-4/7+30ng/mL bFGF,D组:50 ng/mL BMP-4/7+50 ng/mL bFGF,E组:80 ng/mL BMP-4/7+80 ng/mL bFGF)和对照组(不加任何因子),采用绘制生长曲线,甲基噻唑基四唑(MTT)比色法检测细胞增殖活力,碱性磷酸酶(ALP)和降钙素(OC)活性检测法比较各组间差异,观察不同浓度的BMP-4/7和bFGF联合应用对兔BMSCs增殖和成骨活性的影响. 结果传代后第5天对照组个别单核细胞贴壁,呈长梭形;A组细胞增殖,呈旋涡状排列;B组细胞较为密集,部分融合成片;C组细胞呈集落式生长,生长旺盛;D组细胞生长密集,可见明显的钙结节;E组细胞密集,可见细胞性钙结节形成.各组OD值、ALP含量、OC含量随着作用时间的延长而增加,各组不同培养时间的OD值差异均有统计学意义(P<0.01);且C、D、E组均高于A、B组,差异均有统计学意义(P<0.05).C、D、E组内随着作用浓度的增加,细胞增殖及成骨活性增强,呈浓度依赖关系,差异均有统计学意义(P<0.05). 结论合理的联合应用BMP-4/7和bFGF可促进BMSCs细胞增殖,促进成骨活性,两者对BMSCs有明显的协同增强效应.
목적 연구골형태발생단백융합기인-4/7(BMP-4/7)화감성성섬유세포생장인자(bFGF)연합응용대체외배양토골수기질간세포(BMSCs)증식화성골활성적영향.방법 체외배양BMSCs,재제3대세포배양액중가입불동농도적BMP-4/7화bFGF,의거가입불동기인농도조합적불동분위5개실험조(A조:80 ng/mL BMP-4/7,B조:80 ng/mL bFGF,C조:30 ng/mL BMP-4/7+30ng/mL bFGF,D조:50 ng/mL BMP-4/7+50 ng/mL bFGF,E조:80 ng/mL BMP-4/7+80 ng/mL bFGF)화대조조(불가임하인자),채용회제생장곡선,갑기새서기사서(MTT)비색법검측세포증식활력,감성린산매(ALP)화강개소(OC)활성검측법비교각조간차이,관찰불동농도적BMP-4/7화bFGF연합응용대토BMSCs증식화성골활성적영향. 결과 전대후제5천대조조개별단핵세포첩벽,정장사형;A조세포증식,정선와상배렬;B조세포교위밀집,부분융합성편;C조세포정집락식생장,생장왕성;D조세포생장밀집,가견명현적개결절;E조세포밀집,가견세포성개결절형성.각조OD치、ALP함량、OC함량수착작용시간적연장이증가,각조불동배양시간적OD치차이균유통계학의의(P<0.01);차C、D、E조균고우A、B조,차이균유통계학의의(P<0.05).C、D、E조내수착작용농도적증가,세포증식급성골활성증강,정농도의뢰관계,차이균유통계학의의(P<0.05). 결론 합리적연합응용BMP-4/7화bFGF가촉진BMSCs세포증식,촉진성골활성,량자대BMSCs유명현적협동증강효응.
Objective To study the combined effects of basic fibroblast growth factor (bFGF) and fusion bone morpbogenetic gene protein-4/7 (BMP-4/7) on proliferation and osteogenic activity of rabbit bone mesenchymal stem cells (BMSCs) in vitro. Methods BMSCs cells were cultured in vitro. Different concentrations of BMP-4/7 and bFGF were added into the culture of the third passage. Five experimental groups were established according to the concentration of BMP-4/7 and bFGF. Group A: 80 ng/mL BMP-4/7; group B: 80 ng/mL bFGF; group C: 30 ng/mL BMP-4/7 +30 ng/mL bFGF; group D: 50 ng/mL BMP-4/7 +50 ng/mL bFGF; group E: 80 ng/mL BMP-4/7 +80 ng/mL bFGF) . No growth factor was added into the culture fluid in the control. The growth curve drawing, methyl thiazolyl tetrazolium (MTT), alkaline pbosphatase (ALP) and osteocalcin (OC) dynamics were used to compare the differences between the groups and observe the effects of BMP-4/7 plus bFGF on proliferation and osteogenic differentiation of BMSCs. Results Compared with the control group, single BMP-4/7 had promotive effects on the pro-liferation and osteogenic differentiation of BMSCs, while single bFGF contributed to the proliferation of BMSCs significandy hut inhibited the osteogenic differentiation of BMSCs. The combination of BMP-4/7 and bFGF promoted not only the BMSCs proliferation significantly but also the osteogenic differentiation of BMSCs,showing positive correlation with action time and concentration. Conclusion A rational combined use of BMP-4/7 and bFGF can promote the proliferation and osteogenic differentiation of BMSCs and the two factors have significant synergistic effect on BMSCs.